Effect of Gene actA on the Invasion Efficiency of Listeria monocytogenes, as Observed in Healthy and Senescent Intestinal Epithelial Cells

( Jimyeong Ha ),( Hyemin Oh ),( Sejeong Kim ),( Jeeyeon Lee ),( Soomin Lee ),( Heeyoung Lee ),( Yukyung Choi ),( Sung Sil Moon ),( Kyoung-hee Choi ),( Yohan Yoon ) 한국미생물생명공학회(구 한국산업미생물학회) 2018 Journal of microbiology and biotechnology Vol.28 No.1

Listeria monocytogenes can asymptomatically inhabit the human intestine as a commensal bacterium. However, the mechanism by which L. monocytogenes is able to inhabit the intestine without pathogenic symptoms remains unclear. We compared the invasion efficiency of L. monocytogenes strains with the 268- and 385-bp-long actA gene. Clinical strains SMFM-CI-3 and SMFM-CI-6 with 268-bp actA isolated from patients with listeriosis, and strains SMFM-SI-1 and SMFM-SI-2 with the 385-bp gene isolated from carcasses, were used for inoculum preparation. The invasion efficiency of these strains was evaluated using Caco-2 cells (intestinal epithelial cell line), prepared as normal and healthy cells with tightened tight junctions and senescent cells with loose tight junctions that were loosened by adriamycin treatment. The invasion efficiency of L. monocytogenes strains with the 268-bp-long actA gene was 1.1-2.6-times lower than that of the strains with the 385-bp-long gene in normal and healthy cells. However, the invasion efficiency of both types of strains did not differ in senescent cells. Thus, L. monocytogenes strains with the 268-bp-long actA gene can inhabit the intestine asymptomatically as a commensal bacterium, but they may invade the intestinal epithelial cells and cause listeriosis in senescent cells.

Erratum to: Description of Kinetic Behavior of Pathogenic Escherichia coli in Cooked Pig Trotters under Dynamic Storage Conditions Using Mathematical Equations

Ha, Jimyeong,Lee, Jeeyeon,Oh, Hyemin,Kim, Hyun Jung,Choi, Yukyung,Lee, Yewon,Kim, Yujin,Lee, Heeyoung,Kim, Sejeong,Yoon, Yohan Korean Society for Food Science of Animal Resource 2021 한국축산식품학회지 Vol.41 No.2

Kinetic Behavior of Salmonella on Low NaNO₂ Sausages during Aerobic and Vacuum Storage

Ha, Jimyeong,Gwak, Eunji,Oh, Mi-Hwa,Park, Beomyoung,Lee, Jeeyeon,Kim, Sejeong,Lee, Heeyoung,Lee, Soomin,Yoon, Yohan,Choi, Kyoung-Hee Korean Society for Food Science of Animal Resource 2016 한국축산식품학회지 Vol.36 No.2

This study evaluated the growth kinetics of Salmonella spp. in processed meat products formulated with low sodium nitrite (NaNO₂). A 5-strain mixture of Salmonella spp. was inoculated on 25-g samples of sausages formulated with sodium chloride (NaCl) (1.0%, 1.25%, and 1.5%) and NaNO₂ (0 and 10 ppm) followed by aerobic or vacuum storage at 10℃ and 15℃ for up to 816 h or 408 h, respectively. The bacterial cell counts were enumerated on xylose lysine deoxycholate agar, and the modified Gompertz model was fitted to the Salmonella cell counts to calculate the kinetic parameters as a function of NaCl concentration on the growth rate (GR; Log CFU/g/h) and lag phase duration (LPD; h). A linear equation was then fitted to the parameters to evaluate the effect of NaCl concentration on the kinetic parameters. The GR values of Salmonella on sausages were higher (p<0.05) with 10 ppm NaNO₂ concentration than with 0 ppm NaNO₂. The GR values of Salmonella decreased (p<0.05) as NaCl concentration increased, especially at 10℃. This result indicates that 10 ppm NaNO₂ may increase Salmonella growth at low NaCl concentrations, and that NaCl plays an important role in inhibiting Salmonella growth in sausages with low NaNO₂.

Identification of Pork Adulteration in Processed Meat Products Using the Developed Mitochondrial DNA-Based Primers

Jimyeong Ha,Sejeong Kim,Jeeyeon Lee,Soomin Lee,Heeyoung Lee,Yukyung Choi,Hyemin Oh,Yohan Yoon 한국축산식품학회 2017 한국축산식품학회지 Vol.37 No.3

The identification of pork in commercially processed meats is one of the most crucial issues in the food industry because of religious food ethics, medical purposes, and intentional adul-teration to decrease production cost. This study therefore aimed to develop a method for the detection of pork adulteration in meat products using primers specific for pig mitochondrial DNA. Mitochondrial DNA sequences for pig, cattle, chicken, and sheep were obtained from GenBank and aligned. The 294-bp mitochondrial DNA D-loop region was selected as the pig target DNA sequence and appropriate primers were designed using the MUSCLE program. To evaluate primer sensitivity, pork-beef-chicken mixtures were prepared as follows: i) 0% pork-50% beef-50% chicken, ii) 1% pork-49.5% beef-49.5% chicken, iii) 2% pork-49% beef- 49% chicken, iv) 5% pork-47.5% beef-47.5% chicken, v) 10% pork-45% beef-45% chicken, and vi) 100% pork-0% beef-0% chicken. In addition, a total of 35 commercially packaged products, including patties, nuggets, meatballs, and sausages containing processed chicken, beef, or a mixture of various meats, were purchased from commercial markets. The primers developed in our study were able to detect as little as 1% pork in the heat treated pork-beef-chicken mixtures. Of the 35 processed products, three samples were pork positive despite being labeled as beef or chicken only or as a beef-chicken mix. These results indicate that the developed primers could be used to detect pork adulteration in various processed meat prod-ucts for application in safeguarding religious food ethics, detecting allergens, and preventing food adulteration.

Kinetic Behavior of Salmonella on Low NaNO2 Sausages during Aerobic and Vacuum Storage

Jimyeong Ha,Eunji Gwak,Mi Hwa Oh,Beomyoung Park,Jeeyeon Lee,Sejeong Kim,Heeyoung Lee,Soomin Lee,Yohan Yoon 한국축산식품학회 2016 한국축산식품학회지 Vol.36 No.2

Abstract This study evaluated the growth kinetics of Salmonella spp. in processed meat products formulated with low sodium nitrite (NaNO2). A 5-strain mixture of Salmonella spp. was inoculated on 25-g samples of sausages formulated with sodium chloride (NaCl) (1.0%, 1.25%, and 1.5%) and NaNO2 (0 and 10 ppm) followed by aerobic or vacuum storage at 10°C and 15°C for up to 816 h or 408 h, respectively. The bacterial cell counts were enumerated on xylose lysine deoxycholate agar, and the modified Gompertz model was fitted to the Salmonella cell counts to calculate the kinetic parameters as a function of NaCl concentration on the growth rate (GR; Log CFU/g/h) and lag phase duration (LPD; h). A linear equation was then fitted to the parameters to evaluate the effect of NaCl concentration on the kinetic parameters. The GR values of Salmonella on sausages were higher (p<0.05) with 10 ppm NaNO2 concentration than with 0 ppm NaNO2. The GR values of Salmonella decreased (p<0.05) as NaCl concentration increased, especially at 10oC. This result indicates that 10 ppm NaNO2 may increase Salmonella growth at low NaCl concentrations, and that NaCl plays an important role in inhibiting Salmonella growth in sausages with low NaNO2.

Mathematical Models to Describe the Kinetic Behavior of Staphylococcus aureus in Jerky

Jimyeong Ha,Jeeyeon Lee,Soomin Lee,Sejeong Kim,Yukyung Choi,Hyemin Oh,Yujin Kim,Yewon Lee,Yeongeun Seo,Yohan Yoon 한국축산식품학회 2019 한국축산식품학회지 Vol.39 No.3

The objective of this study was to develop mathematical models for describing the kinetic behavior of Staphylococcus aureus (S. aureus) in seasoned beef jerky. Seasoned beef jerky was cut into 10-g pieces. Next, 0.1 mL of S. aureus ATCC13565 was inoculated into the samples to obtain 3 Log CFU/g, and the samples were stored aerobically at 10°C, 20°C, 25°C, 30°C, and 35°C for 600 h. S. aureus cell counts were enumerated on Baird Parker agar during storage. To develop a primary model, the Weibull model was fitted to the cell count data to calculate Delta (required time for the first decimal reduction) and ρ (shape of curves). For secondary modeling, a polynomial model was fitted to the Delta values as a function of storage temperature. To evaluate the accuracy of the model prediction, the root mean square error (RMSE) was calculated by comparing the predicted data with the observed data. The surviving S. aureus cell counts were decreased at all storage temperatures. The Delta values were longer at 10°C, 20°C, and 25°C than at 30°C and 35°C. The secondary model well-described the temperature effect on Delta with an R2 value of 0.920. In validation analysis, RMSE values of 0.325 suggested that the model performance was appropriate. S. aureus in beef jerky survives for a long period at low storage temperatures and that the model developed in this study is useful for describing the kinetic behavior of S. aureus in seasoned beef jerky.

Description of Kinetic Behavior of Pathogenic Escherichia coli in Cooked Pig Trotters under Dynamic Storage Conditions Using Mathematical Equations

Jimyeong Ha,Jeeyeon Lee,Hyemin Oh,Hyun Jung Kim,Yukyung Choi,Yewon Lee,Yujin Kim,Heeyoung Lee,Sejeong Kim,Yohan Yoon 한국축산식품학회 2020 한국축산식품학회지 Vol.40 No.6

A dynamic model was developed to predict the Escherichia coli cell counts in pig trotters at changing temperatures. Five-strain mixture of pathogenic E. coli at 4 Log CFU/g were inoculated to cooked pig trotter samples. The samples were stored at 10℃, 20℃, and 25℃. The cell count data was analyzed with the Baranyi model to compute the maximum specific growth rate (μmax) (Log CFU/g/h) and lag phase duration (LPD) (h). The kinetic parameters were analyzed using a polynomial equation, and a dynamic model was developed using the kinetic models. The model performance was evaluated using the accuracy factor (Af), bias factor (Bf), and root mean square error (RMSE). E. coli cell counts increased (p<0.05) in pig trotter samples at all storage temperatures (10℃-25℃). LPD decreased (p<0.05) and μmax increased (p<0.05) as storage temperature increased. In addition, the value of h0 was similar at 10℃ and 20℃, implying that the physiological state was similar between 10℃ and 20℃. The secondary models used were appropriate to evaluate the effect of storage temperature on LPD and μmax. The developed kinetic models showed good performance with RMSE of 0.618, Bf of 1.02, and Af of 1.08. Also, performance of the dynamic model was appropriate. Thus, the developed dynamic model in this study can be applied to describe the kinetic behavior of E. coli in cooked pig trotters during storage.

Quantitative Microbial Risk Assessment for Campylobacter spp. on Ham in Korea

Lee, Jeeyeon,Ha, Jimyeong,Kim, Sejeong,Lee, Heeyoung,Lee, Soomin,Yoon, Yohan Korean Society for Food Science of Animal Resource 2015 한국축산식품학회지 Vol.35 No.5

The objective of this study was to evaluate the risk of illness from Campylobacter spp. on ham. To identify the hazards of Campylobacter spp. on ham, the general characteristics and microbial criteria for Campylobacter spp., and campylobacteriosis outbreaks were investigated. In the exposure assessment, the prevalence of Campylobacter spp. on ham was evaluated, and the probabilistic distributions for the temperature of ham surfaces in retail markets and home refrigerators were prepared. In addition, the raw data from the Korea National Health and Nutrition Examination Survey (KNHNES) 2012 were used to estimate the consumption amount and frequency of ham. In the hazard characterization, the Beta-Poisson model for Campylobacter spp. infection was used. For risk characterization, a simulation model was developed using the collected data, and the risk of Campylobacter spp. on ham was estimated with @RISK. The Campylobacter spp. cell counts on ham samples were below the detection limit (<0.70 Log CFU/g). The daily consumption of ham was 23.93 g per person, and the consumption frequency was 11.57%. The simulated mean value of the initial contamination level of Campylobacter spp. on ham was −3.95 Log CFU/g, and the mean value of ham for probable risk per person per day was 2.20×10⁻¹². It is considered that the risk of foodborne illness for Campylobacter spp. was low. Furthermore, these results indicate that the microbial risk assessment of Campylobacter spp. in this study should be useful in providing scientific evidence to set up the criteria of Campylobacter spp..

The Correlation between NaCl Adaptation and Heat Sensitivity of Listeria monocytogenes, a Foodborne Pathogen through Fresh and Processed Meat

Lee, Jeeyeon,Ha, Jimyeong,Kim, Sejeong,Lee, Soomin,Lee, Heeyoung,Yoon, Yohan,Choi, Kyoung-Hee Korean Society for Food Science of Animal Resource 2016 한국축산식품학회지 Vol.36 No.4

This study examined the relationship between NaCl sensitivity and stress response of Listeria monocytogenes. Nine strains of L. monocytogenes (NCCP10805, NCCP10806, NCCP10807, NCCP10808, NCCP10809, NCCP10810, NCCP10811, NCCP10920 and NCCP 10943) were exposed to 0%, 1%, 2% and 4% NaCl, and then incubated at 60℃ for 60 min to select strains that were heat-sensitized (HS) and non-sensitized (NS) by NaCl exposure. After heat challenge, L. monocytogenes strains were categorized as HS (NCCP 10805, NCCP10806, NCCP10807, NCCP10810, NCCP10811 and NCCP10920) or NS (NCCP10808, NCCP10809 and NCCP10943). Total mRNA was extracted from a HS strain (NCCP10811) and two NS strains (NCCP10808 and NCCP10809), and then cDNA was prepared to analyze the expression of genes (inlA, inlB, opuC, betL, gbuB, osmC and ctc) that may be altered in response to NaCl stress, by qRT-PCR. The expression levels of two invasion-related genes (inlA and inlB) and two stress response genes (opuC and ctc) were increased (p<0.05) in NS strains after NaCl exposure in an NaCl concentration-dependent manner. However, only betL expression was increased (p<0.05) in the HS strains. These results indicate that the effect of NaCl on heat sensitization of L. monocytogenes is strain dependent and that opuC and ctc may prevent NS L. monocytogenes strains from being heat sensitized by NaCl. Moreover, NaCl also increases the expression of invasion-related genes (inlA and inlB).

High Prevalence of Listeria monocytogenes in Smoked Duck: Antibiotic and Heat Resistance, Virulence, and Genetics of the Isolates

Park, Eunyoung,Ha, Jimyeong,Oh, Hyemin,Kim, Sejeong,Choi, Yukyung,Lee, Yewon,Kim, Yujin,Seo, Yeongeun,Kang, Joohyun,Yoon, Yohan Korean Society for Food Science of Animal Resource 2021 한국축산식품학회지 Vol.41 No.2

This study aimed at determining the genetic and virulence characteristics of the Listeria monocytogenes from smoked ducks. L. monocytogenes was isolated by plating, and the isolated colonies were identified by PCR. All the obtained seven L. monocytogenes isolates possessed the virulence genes (inlA, inlB, plcB, and hlyA) and a 385 bp actA amplicon. The L. monocytogenes isolates (SMFM2018 SD 1-1, SMFM 2018 SD 4-1, SMFM 2018 SD 4-2, SMFM 2018 SD 5-2, SMFM 2018 SD 5-3, SMFM 2018 SD 6-2, and SMFM 2018 SD 7-1) were inoculated in tryptic soy broth (TSB) containing 0.6% yeast extract at 60℃, followed by cell counting on tryptic soy agar (TSA) containing 0.6% yeast extract at 0, 2, 5, 8, and 10 min. We identified five heat resistant isolates compared to the standard strain (L. monocytogenes ATCC13932), among which three exhibited the serotype 1/2b and D-values of 5.41, 6.48, and 6.71, respectively at 60℃. The optical densities of the cultures were regulated to a 0.5 McFarland standard to assess resistance against nine antibiotics after an incubation at 30℃ for 24 h. All isolates were penicillin G resistant, possessing the virulence genes (inlA, inlB, plcB, and hlyA) and the 385-bp actA amplicon, moreover, three isolates showed clindamycin resistance. In conclusion, this study allowed us to characterize L. monocytogenes isolates from smoked ducks, exhibiting clindamycin and penicillin G resistance, along with the 385-bp actA amplicon, representing higher invasion efficiency than the 268-bp actA, and the higher heat resistance serotype 1/2b.